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. 2021 Jul 27;12:4559. doi: 10.1038/s41467-021-24876-1

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Percentage of (a) GFP⁺ cells within the human graft (hCD45⁺), (b) human engrafted cells in total BM and (c) myeloid (CD33⁺) and (d) lymphoid B cells (CD19⁺) within the human graft at euthanasia, in the BM of mice transplanted with CTRL (wtBRAF or GFP only, blue, n = 31) and BRAF^V600E (red, n = 25) cells. Data are presented as mean +/− SD. Statistical test: Two-tailed Mann−Whitney. e Percentage of myeloid GFP⁺ (red, dashed green) and myeloid GFP⁻ (red), B (CD19⁺) cells GFP⁻ (blue) and B cells GFP⁺ (blue, dashed green) within total BM from mice transplanted with CTRL or BRAF^V600E HSPCs. Values on x-axis were ordered by transduction level. Percentage of GFP⁺ and GFP⁻ (f) B cells or (g) myeloid cells within total BM from mice transplanted with CTRL (blue, n = 31 for left panel, n = 22 for mid and right panel) and BRAF^V600E-expressing (red, n = 25) HSPCs. Data are represented as mean values +/− SD. Statistical test: two-tailed Mann−Whitney. h Percentage of GFP⁺ and GFP⁻ CD19⁺ cells with respect to CD45⁺/GFP⁺ in each mouse. Exponential regression analysis. The goodness of fit measured as R². i Time-course of infiltration grade in organs from mice of the wtBRAF (blue) and BRAF^V600E (red) groups; n = 3 for each time point. Data are presented as mean +/− SD. Spleen: dashed line, circles; liver: dashed line, squares; lung: dashed line, triangles; BM: continuous line, dots. j Time-course analyses of the percentage of GFP⁺ and GFP⁻ myeloid and B cells in BM of mice transplanted with wtBRAF or BRAF^V600E HSPCs; n = 3 each time point. Data are presented as mean +/− SD, see (e) for color code. k–p Percentage of human HSPCs subpopulations within GFP⁻ and GFP⁺ cells in BM cells harvested from mice at 9 and 14 days after transplant with CTRL (untransduced, blue, n = 2 for each time point), wtBRAF (blue, n = 5 for each time point) and BRAF^V600E (red, n = 5 for each time point) HSPCs. Data are presented as mean +/− SD (HSPC hematopoietic stem and progenitor cells; HSC hematopoietic stem cells; MLP multi-lymphoid progenitors; PreBNK B and NK cell progenitors; CMP common-myeloid progenitors; GMP granulocyte/monocyte progenitors). Statistical analysis between groups by two-tailed Mann−Whitney test. ns non-significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.